L'endocytose dépendante de la clathrine (EDC), c’est-à-dire la formation de vésicules recouvertes de clathrine (VRC) à partir de la membrane plasmique, est un processus essentiel dans les cellules eucaryotes. Au cours de l’EDC, la GTPase dynamine est recrutée au cou de la VRC naissante où elle s'oligomérise en hélice. Les changements de conformation induits par l'hydrolyse du GTP catalysent la scission du cou vésiculaire. Ce processus a été étudié en détail par reconstitution in vitro sur des tubules membranaires, mais il doit être établi dans des cellules vivantes, où les interactions de la dynamine avec d'autres protéines comme l'amphiphysine sont critiques. L'imagerie TIRF (Total Internal Reflection Fluorescence) avec le protocole pH pulsé (ppH) sur cellules vivantes permet la détection de la formation de VRC avec une résolution spatiale (~100 nm) et temporelle (2 s) élevée. Ce protocole a révélé que la dynamine présente un recrutement biphasique aux puits recouverts de clathrine (PRC) en maturation avec un pic au moment de la scission mais les paramètres de son recrutement dans les cellules vivantes restent peu clairs. Pour déterminer ces paramètres, j’ai utilisé des techniques d’imagerie sur cellules vivantes pour étudier le recrutement de la dynamine à l’échelle globale et à l’échelle de la molécule unique lors de perturbations aiguës de sa fonction. Mes résultats de thèse ont montré que la dynamine est recrutée à la membrane plasmique, diffuse à l'extérieur des PRC et y est transitoirement piégée. De plus, j’ai déterminé avec des dynamines mutées (1) que le domaine PRD de la dynamine est crucial pour son recrutement aux PRC ; (2) que le domaine PH est important pour la scission vésiculaire mais par pour son recrutement aux PRC ou à la membrane plasmique. Enfin, j’ai observé que la dynamine s'échange en permanence avec un pool extra-PRC, ce qui permettrait son recrutement ultérieur par l'ajout de nouveaux sites de liaison et sa capacité à rétrécir le cou des vésicules suite à l’hydrolyse du GTP. En conclusion, ces données suggèrent qu’aux PRC, les molécules de dynamine (1) sont constamment échangées ; (2) diffusent à des taux similaires tout au long du processus de formation, maturation et scission des vésicules; et (3) l'activité GTPase de la dynamine contribue à la maturation et à la scission des VRC. / Clathrin-mediated endocytosis (CME), the formation of clathrin-coated vesicles (CCV) from the plasma membrane, is an essential process in eukaryotic cells. During CME, the GTPase dynamin is recruited to the neck of nascent CCV where it oligomerizes into helical filaments. Conformational changes induced by the hydrolysis of GTP catalyze the scission of the vesicle neck. This process has been studied in detail with in vitro reconstitution on membrane tubules but it needs to be established in living cells, where interactions between dynamin and other proteins such as amphiphysin are critical. Live cell total internal reflection fluorescence (TIRF) imaging with the pulsed pH (ppH) assay allows the detection of CCV formation with high spatial (~100 nm) and temporal (2 s) resolutions. It has revealed that dynamin is recruited to maturing clathrin-coated pits (CCP) in two phases with a peak at the time of scission but the parameters of its recruitment in living cells remain unclear. To determine these parameters, we have performed live cell imaging of dynamin recruitment at collective and single molecule levels during acute perturbations of its function. My PhD results showed that dynamin is recruited to the plasma membrane, diffuses outside of CCP and is trapped at CCP. Furthermore, we determined with mutated dynamins that (1) the PRD domain of dynamin is crucial for its recruitment at CCP; (2) the PH domain is important for vesicular scission but not for recruitment to CCP or to the plasma membrane. Finally, I observed that dynamin exchanges with an extra-CCP pool at all times: this would allow for its further recruitment by addition of new binding sites and its ability to narrow the vesicle neck after GTP hydrolysis. Altogether, these data suggest that in CCP dynamin molecules (1) are constantly exchanged; (2) diffuse at similar rates throughout the entire process of vesicle formation, from maturation until scission; and (3) that dynamin’s GTPase activity contributes to CCP maturation and scission.

No description available.

[pt] Nesta dissertação, é apresentado um procedimento estatístico para a estimação de perfis de curvas de carga por aparelhos para consumidores residenciais atendidos na BT (Baixa Tensão). O modelo desenvolvido utiliza informações de duas fontes distintas: PPHs (Pesquisas de Posses e Hábitos de Aparelhos Elétricos) e Medições de consumo com medidores eletrônicos com memória de massa que gravam leituras de consumo por aparelhos a cada 15 minutos. Através das PPHs desenvolvidas por Procel/Eletrobrás em 1996, obtêm-se um perfil aproximado da curva de carga do domicilio por aparelho. Entretanto, esta curva tende a ser imprecisa, pois é obtida por declarações de uso dos aparelhos. Para reduzir esta imprecisão, foi montado um experimento onde uma sub-amostra de domicílios auditados pelas PPHs, teve os principais aparelhos da residência medidos por medidores eletrônicos em intervalos de 15 minutos. A partir das curvas de utilização destes aparelhos, obtidas pelos dois procedimentos, utilizou-se um modelo estatístico de regressão linear para estimar coeficientes de ajustes para correção das curvas declaradas para cada hora do dia, para os principais eletrodomésticos do domicílio. Os resultados foram aplicados a duas distribuidoras do grupo ENDESA: AMPLA e COELCE. / [en] In this thesis, it is presented a statistical based model that allows the estimation of the load shape curve for appliances (end uses) for residential consumers that belong to low voltage group (BT clients), using information from two sources: PPHs (Energy Audit on Ownership and Usage of Electrical Appliances) and household measurements through specific devices that provide not only the total consumption (15 minute intervals) as well as the breakdown of this consumption for each device (end use) existing in the household. Through energy audits (PPHs) developed by Procel/Eletrobrás in 1996, one has a rough idea of the load shape curves by appliance. However, the curves obtained this way tend to be rather imprecise, as they are obtained by the consumer information of usage of the equipments on the surveys, which tend to be rather vague. In order to reduce such imprecision, an experiment was set where a subsample of the original survey sample is selected and the main appliances consumption measured by electronic meters at intervals of 15 minutes. The end use load shape obtained by these meters are then compared with the corresponding curve obtained by usage declaration. Using the linear regression model, the correction coefficients of the declarations are obtained for each hour of the day, for the main appliances. The results were applied to two distribution utilities of the ENDESA group: AMPLA and COELCE.

A Thesis submitted to The University of Arizona College of Medicine - Phoenix in partial fulfillment of the requirements for the Degree of Doctor of Medicine. / Primary pulmonary hypertension (PPH) has an extremely poor prognosis with a mean survival time of 2‐3 years from time of diagnosis. Hemodynamically, PPH is defined with a mPAP of ≥ 25 mm Hg. Currently, RHC is the gold standard for measuring the arterial pressures and diagnosing PPH; however, it is an incredibly invasive procedure. Our study will show whether CT angiography can be considered as a non‐invasive alternative for diagnosing PPH. Studies in the past have shown CT measurements of the MPAD and MPAD/AAD ratio having strong correlations with PPH. In addition to those measurements, we want to show if other CT parameters also have a correlation with PPH. Some of these novel measurements include the interventricular septal deviation and the Elizabeth Taylor sign. The interventricular septum is normally bowing to the right in a non‐pathological state. If it is straight or bowing to the left, this will indicate increased right ventricular pressures which would be indicative of PPH. Straight will indicate increased RV pressures, and bowing to the left will be considered markedly increased RV pressures. The Elizabeth Taylor sign is the ratio of the diameter of the segmental bronchi and its corresponding artery. We will hypothesize that the artery will be much larger than the bronchi in patients with PPH. Other measurements will include the left and right pulmonary arteries. This study is a retrospective review of subjects who underwent an otherwise unremarkable CT pulmonary artery angiogram. Subjects with pulmonary embolism or other acute pulmonary diseases are excluded. For each subject, the following CT findings are obtained: main pulmonary artery diameter (mPAD), ratio of mPAD to ascending aorta, right and left pulmonary artery diameters, ratio of segmental pulmonary artery to corresponding bronchus, and interventricular septal displacement. Straightening of the interventricular septum qualifies as increased right ventricular septal pressure and right‐to‐left bowing of the septum qualifies as a marked increase. Mean pulmonary artery pressure measured on any prior/subsequent RHC or echocardiogram within 3 months of the CT is recorded. Any past medical history of connective tissue disease is noted. Descriptive data are calculated and correlations are done to assess for presence and strength of associations among variables. Data from 484 subjects are collected. Incidence rate of pulmonary hypertension isv13% (n=63). 52% (n=33) of the subjects with pulmonary hypertension are female with an average age of 55 years. mPA diameter (p<0.001), mPA:AA ratio (p<0.001), right (p<0.001) and left pulmonary artery (p=0.004) diameters are predictors of pulmonary hypertension. sPA:B ratio (p=0.08) and interventricular septal displacement (p=0.96) are not predictive of pulmonary hypertension. This study supports an association of mPA diameter, mPA:AA ratio, right and left pulmonary artery diameters with pulmonary hypertension diagnosed by RHC or echocardiogram. Prospective research is warranted to confirm and establish threshold values for each variable. Currently, an invasive RHC remains the most accurate method of diagnosis. Correlating CT findings with pulmonary hypertension would allow clinicians to use CT as a noninvasive screening tool.

No description available.

L’hypertension artérielle pulmonaire (HTAP) correspond à une augmentation des résistances artérielles pulmonaires, avec dans les formes les plus graves, une défaillance cardiaque droite. L’HTAP persistante du nouveau-né (HTAPP) est estimée à 2/100 naissances en France. L’adaptation cardio-respiratoire à la naissance implique le déclenchement simultané de plusieurs phénomènes non complètements compris. Notre travail avait pour but d’approfondir les connaissances sur la régulation pulmonaire périnatale et d’envisager de nouvelles possibilités thérapeutiques. Ainsi, nous avons mis en évidence in vivo une vasoconstriction pulmonaires chez le fœtus lors du tabagisme passif maternel, par blocage de la voie du NO. De même, nous avons pu montrer l’effet vasoconstricteur de l’apeline sur les artères pulmonaires de fœtus de brebis. Cet effet semble dose dépendant, inhibé par l’action des inhibiteurs calciques. Au contraire nous avons mis en évidence un effet vasodilatateur de la Déhydroépiandrostérone (DHEA). Cette action est médiée par la voie du NO. Parallèlement, nous avons montré que les acides gras poly insaturés &#969; 3 (AGPI&#969;3) entrainaient une vasodilatation pulmonaire, se prolongeant au delà d’une heure après l’arrêt de la perfusion. Cet effet est médié par l’ouverture des canaux potassiques et indépendant de la voie du NO. Parmi les AGPI&#969;3, nous avons établi que l’acide eicosapentaénoïque (EPA) qui induit cette réponse sans effet délétère sur la circulation systémique, ni l’oxygénation tissulaire. Enfin, nous avons établi que l’occlusion trachéale (OT) anténatale, traitement proposé pour certaine hypoplasie pulmonaire malformative, n’altère pas le débit pulmonaire, en favorisant la dilatation pulmonaire. Mais en cas d’OT prolongée, ces effets sont en partie masqués par les effets mécaniques de la pression intraluminale. Ainsi grâce à ces travaux, nous avons avancé dans la compréhension de l’adaptation de la circulation pulmonaire de la vie intra à la vie extra utérine. Ils permettent aussi de proposer de nouvelles thérapeutiques comme la supplémentation en AGPI&#969; 3 des femmes attendant un enfant à risque d’HTAP, et d’envisager de nouvelles voies de recherche thérapeutique comme la voie de l’apeline. / Persistent pulmonary hypertension (PPH) corresponds to an increase in pulmonary arterial resistance, with in the most severe forms, right heart failure. The persistent pulmonary hypertension of the newborn (PPHN) is estimated at 2/100 births in France. The cardiorespiratory adaptation at birth involves the simultaneous triggering of several phenomena including non completions. Our work aimed to increase knowledge on the regulation of perinatal pulmonary and consider new therapeutic possibilities. Thus, we have demonstrated in vivo pulmonary vasoconstriction in the fetus when maternal passive smoking, by blocking the NO pathway. Similarly, we demonstrated the vasoconstrictor effect of apelin infusion on the pulmonary arteries of fetal sheep. This effect appears dose dependent, inhibited by the action of calcium channel blockers. Instead we have shown a vasodilatory effect of dehydroepiandrosterone (DHEA). This action is mediated by the NO pathway. In parallel, we have shown that polyunsaturated fatty acids &#969; 3 (AGPI&#969;3) would cause pulmonary vasodilation, extending beyond an hour after stopping the infusion. This effect is mediated by the opening of potassium channels and independent of the NO pathway. Among AGPI&#969;3, we found that eicosapentaenoic acid (EPA) that induces this response without deleterious effect on the systemic circulation or tissue oxygenation. Finally, we established that the antenatal tracheal occlusion (TO), treatment proposed for some malformative pulmonary hypoplasia, does not alter pulmonary blood flow, promoting lung expansion. But in case of prolonged OT, these effects are partly masked by the mechanical effects of intraluminal pressure. And through this work, we have made progress in understanding the adaptation of the pulmonary circulation of the intra to extrauterine life. They also suggest new therapeutic, like supplementation as AGPI&#969; for women expecting a child at risk for PAH, and to consider new possibility of therapeutic research like the way of apelin.

[pt] O objetivo desta dissertação é desenvolver modelos matemáticos que permitam estimar o tempo médio dos banhos com a utilização de chuveiros elétricos e a curva de carga desses aparelhos, considerando as informações das Pesquisas de Posses e Hábitos de Consumo (PPH) e medições realizadas com o auxílio de medidores eletrônicos com memória de massa, em residências com chuveiros elétricos. A motivação do estudo advém de uma exigência da ANEEL que determina que as distribuidoras de energia elétrica realizem a cada 2 (dois) ciclos de revisão tarifária a PPH em suas unidades consumidoras. Os métodos empregados foram: estatística descritiva (para a obtenção do tempo médio de banho); aplicação da regressão linear e de redes neurais (para corrigir a curva de carga horária obtida com a PPH, com base nos dados das medições). Os resultados foram promissores, pois o tempo médio de banho se encontra próximo às estimativas do PROCEL (que são de 8 (oito) a 10 (dez) minutos) e a curva de carga estimada se encontra próxima à da medição, sendo esta última o consumo real. Conclui-se que a abordagem desta dissertação resultou em melhorias na estimativa dos coeficientes de ajustes e que o método de redes neurais foi relativamente melhor que o método de regressão linear simples. / [en] The aim of this dissertation is to develop mathematical models that would allow the estimation of the average time of baths using electric showers and the load shape curves for these devices, obtained from two sources: the information of Electrical Appliances Ownership Survey and measurements of electric shower usage in households carried out with electronic meters with storage capacity. The motivation stems from a requirement of ANEEL that determines that the electric energy distributors periodically should hold a PPH in their consumer units. Concerning the average time of shower baths, the last PPH survey conducted by PROCEL in 2005 estimated this time between 8 (eight) and 10 (ten) minutes. The methods employed in this work were: descriptive statistics (for obtaining the average bath time); application of linear regression and neural networks (to estimate the correction factors to approximate the load shape curves obtained by PPH to those obtained by measurements). The obtained results are rather promising due to the following reasons: the average time of bath is next to the estimates of PROCEL and the corrected load shape curve estimated is quite close to the measured curve, the latter being the actual consumption. This approach has resulted in improvements in the estimation of the coefficients of adjustments and the method of neural networks was relatively better than the simple linear regression method.